Posted by
Prodanov, D. (FYS) on
Sep 21, 2005; 9:57am
URL: http://imagej.273.s1.nabble.com/background-removal-in-fluorescence-tp3704817p3704818.html
Dear Martin,
Actually, what the background subtraction does is to use the "rolling ball"
Image is sampled in a ratio and then degraded by a Structuring Element.
then the degraded image is subtracted from the initial image (i.e. "top hat" transform).
Anyway, I think you're on the right track. May be you should do
stack averaging? Or may be you should track the intensity of a praticular structure
and readjust the histograms according to it.
best regards
Dimiter
Date: Tue, 20 Sep 2005 13:11:52 -0500
From: Martin Wessendorf <
[hidden email]>
Subject: Re: background removal in fluorescence
Dear ImageJ-ers--
I realize that I didn't articulate the problem particularly well. --We
want to make some montages of images. Our illumination is not perfectly
even and so parts of the field are illuminated more brightly than
others. Thus when we try to make the montage, there are abrupt
inconsistencies in the lighting across it.
I had thought that the following *should* work:
1) Take an image of a uniform fluorescent field (image A)
2) Take an image of the specimen (image B)
3) Divide B by A; multiply by the mean intensity of A.
(--I think that this is what Wayne's plugin does.)
However, when we use that method, the inconsistencies of the
illumination are greatly accentuated rather than being reduced. My
guess is that this is due to non-linearity of response in our CCD camera.
I'd be delighted if anyone has a work-around for this problem!
Martin